南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (08): 964-.doi: 10.12122/j.issn.1673-4254.2019.08.14

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脑络欣通对神经干细胞增殖分化及β-tubulin III/GFAP的影响

何玲，石晓倩，徐伟，谭辉，王键   

  • 出版日期:2019-08-20 发布日期:2019-08-20

Effect of Naoluo Xintong on proliferation and differentiation of neural stem cells and β-tubulin III/GFAP

  • Online:2019-08-20 Published:2019-08-20

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摘要： 目的观察脑络欣通对β-tubulinⅢ/GFAP及神经干细胞（NSCs）增殖分化的影响。方法将NSCs细胞株分为空白模型 （MC）组、10%脑络欣通含药血清（NLXT组）、10%脑络欣通含药血清+抑制剂Y27632组（Y-27632组）。采用MTT法检测NSCs 细胞活性，Transwell小室观察NSCs迁移，免疫印迹法观察β-TubulinⅢ、GFAP、MAP-2蛋白表达。免疫荧光标记DCX、NEUN、 β-TubulinⅢ表达。结果与MC组比较，1 d和3 d NLXT组、Y-27632组迁移细胞数增多（P<0.05）；7 d后NLXT组、Y-27632组细 胞存活率及迁移细胞数显著增多（P<0.01）。与MC组比较，第3 天、第7 天NLXT组和Y-27632组β-tubulinⅢ、MAP2、GFAP蛋 白表达显著升高（P<0.01或P<0.05）。NLXT组、Y-27632组脑组织β-tubulinⅢ/GFAP、BrdU/DCX、BrdU/NEUN标记细胞数目较 MC组明显增多。结论脑络欣通通过调节β-tubulinⅢ/GFAP促进NSCs增殖、分化。

Abstract: Objective To observe the effects of Naoluo Xintong on the expression of β-tubulin III and glial fibrillary acidic protein (GFAP) and the proliferation and differentiation of murine neural stem cells (NSCs) in vitro. Methods An immortalized murine NSC line was divided into model control (MC) group, 10% Naoluo Xintong drug-containing serum group (NLXT group), and 10% Naoluoxintong drug-containing serum with inhibitor Y27632 group (Y-27632 group) with corresponding treatments. The activity of the NSCs was detected after the treatments using MTT assay, and the migration of the cells was observed with Transwell assay. The expressions of β-tubulin III, GFAP and MAP-2 proteins in the cells were detected with immunoblotting, and the expressions of DCX, NEUN, and β-tubulin III were also detected with immunofluorescence assay. Results Compared with that in MC group, the number of migrated cells in NLXT group and Y-27632 group increased significantly at 1 day and 3 days after induction (P<0.05). The survival rate and the number of migrated cells in NLXT group and Y-27632 group increased significantly on day 7 (P<0.01). Compared with those in MC group, the expressions of β-tubulin III, MAP2 and GFAP protein in NLXT group and Y-27632 group were significantly increased on days 3 (P<0.01) and 7 (P<0.05). The numbers of β-tubulinIII/ GFAP, BrdU/DCX, and BrdU/NEUN labeled cells in the NLXT group and Y-27632 group were significantly greater than those in the MC group. Conclusion Naoluo Xintong promotes the proliferation and differentiation of murine NSCs in vitro by regulating the expressions of β-tubulinIII/GFAP.